福建省大豆炭疽病病原菌的分离与鉴定

为明确引起福建省大豆炭疽病的病原菌种类,2018—2019年从福建省福州、三明、莆田、泉州和南平市采集具有大豆炭疽病症状的大豆豆荚,采用组织分离法分离病原菌,结合形态学特征和多基因系统发育分析对病原菌进行鉴定。结果表明,从采集的豆荚样本中共分离获得152株炭疽菌菌株,经形态学初步鉴定分属于3种类型,从3种类型的病原菌中分别选取3株代表菌株进行多基因系统发育分析,这9株代表菌株分别与Colletotrichum plurivorum、平头炭疽菌C.truncatum和胶孢炭疽菌C.gloeosporioides聚在一起。形态学和多基因系统发育分析结果表明,引起福建省大豆炭疽病的病原菌有3种,分别为C.plurivorum、平头炭疽菌和胶孢炭疽菌,其占比分别为10.53%、50.00%、39.47%。致病性测定结果表明,C.plurivorum、平头炭疽菌和胶孢炭疽菌对大豆均有致病作用,但不同的菌株致病力存在差异。表明平头炭疽菌是引起福建省大豆炭疽病的主要病原菌,而C.plurivorum为引起我国大豆炭疽病的新纪录种。     

浙江省鲜食大豆炭疽病病原分离及抗性鉴定

 炭疽病是我国南方鲜食大豆最重要的病害之一。本试验通过组织分离法,在浙江省鲜食大豆主栽区炭疽病病荚中分离得到9株疑似炭疽病菌分离物。根据分离物的形态特征、基于ITS-GAPDH-CHS-1-TUB2基因序列的系统发育分析、分离物的致病性将这些分离物鉴定为平头炭疽菌(Colletotrichum truncatum)。对51份栽培大豆品种鼓粒初期豆荚离体喷雾接种强致病力分离物CT5,这些品种表现出明显抗/感差异,其病斑大小被用于大豆平头炭疽菌抗病性分级标准。在供试的这些品种中,发现了3份高抗品种,9份抗病品种,20份中抗品种。本研究明确了浙江省鲜食大豆炭疽病病原,提供了部分大豆品种资源抗病性信息,为深入研究该病发生规律、防治、抗病机制及培育抗病新品种等奠定基础。     

海南省温郁金炭疽病的病原鉴定

温郁金炭疽病是一种重要的真菌性病害,该病主要危害叶片,初期产生水渍状的淡黄色病斑,后逐渐扩大成圆形、椭圆形或不规则形的灰褐色病斑,上面有黑色小点,病斑外面环绕一层黄色晕圈,严重时多个病斑联合造成叶片卷缩和枯死,2016年我们从海南省的温郁金种植基地采集炭疽病样本,经病原菌的分离纯化、柯赫氏法则验证、病原菌的形态特征观察及多基因谱系分析方法,确定其病原菌为郁金炭疽菌Colletotrichum curcumae(Syd.)E.J.ButlerBisby。     

毛豆炭疽病的发生与防治

毛豆又名枝豆 ,是一种菜用大豆 ,是龙海市创汇农业的主要经济作物之一 ,年种植面积 4 0 0 0～ 50 0 0ｈｍ2 。近几年来 ,毛豆炭疽病 (黑斑荚 )发生较为严重 ,严重田块病荚率达 85%以上 ,一旦发生黑斑荚就失去商品价值 ,经济损失严重。为此 ,笔者对该病的防治技术进行了研究探讨。现将研究情况初报如下 :1　病原与症状此病是由大豆炭疽病菌 (Ｃｏｕｔｏｔｒｉｃｈｕｍｇｌｙｃｉｎｅｓ)真菌侵染引起的。主要为害豆荚。起初在幼荚表皮产生褪绿色斑点 ,以后逐渐扩大 ,形成不规则形褐色斑 ,在结荚后期及气候潮湿的条件下 ,在病组织上密生黑色小…     

菜豆炭疽病的防治

菜豆炭疽病是菜豆常见的病害 ,该病危害菜豆的叶、茎、荚、种子 ,特别是豆荚受害后 ,严重影响菜豆的质量 ,使菜豆失去应有的商品价值。１受害症状菜豆炭疽病危害幼苗时 ,幼苗子叶上出现红褐色至黑褐色圆形斑 ,凹陷成溃疡状 ,叶上染病多发生在叶背的叶脉上 ,为黑褐色多角形小斑 ;茎上的病斑一般长圆形、稍凹陷 ;豆荚感病 ,初为褐色小点 ,后扩大成近圆形、稍凹陷黑褐色、边缘有深粉红色晕圈的病斑 ;种子上的病斑为黑褐色或黄褐色小斑。２防治方法２．１从无病的豆荚上采种播种前用种子重量的０．２%的５０ %多菌灵粉剂拌种 ,亦可用甲醛２００…     

辽宁省北镇市葡萄叶斑病的病原鉴定

近年来辽宁省北镇市出现一种葡萄叶斑病,症状表现为圆形或椭圆形小黑斑,病斑周围褪绿变黄,为明确引起该病的病原菌,利用组织分离法对病样进行分离纯化,采用多基因系统发育学分析(ITS、GAPDH、CHS-1、ACT、TUB2基因)和形态学特征观察对纯化后的菌株进行鉴定,并结合柯赫氏法则对其进行验证。结果确定病原菌为平头炭疽菌Colletotrichum truncatum.     

烟草弯孢炭疽病病原鉴定及生物学特性

烟草是我国重要的经济作物,烟草病害已成为制约烟草生产的重要因素。烟草弯孢炭疽病是本实验室在山东莒县发现的一种烟草病害,主要危害烟草叶片,病斑圆形至椭圆形,多个病斑连接后成不规则形大斑,后干枯、脱落,造成叶片穿孔,严重时仅剩烟筋,丧失其经济价值。该病病原为产新月形分生孢子的炭疽菌,到目前为止国内尚未见该病害的相关报道。本研究对该病原菌进行了鉴定,并探讨其生物学特性。     

阳春砂仁炭疽病(叶枯病)的病原鉴定

阳春砂仁为姜科多年生草本植物,是我国著名的中药材,以果入药,经济价值较高.近因炭疽病发生严重,砂仁生产受到很大威胁.此病主要在叶片发生,从叶尖向下变成灰褐色枯死,或在叶片上有圆形或不规则形状的褐色或灰褐色斑,病斑稍凹陷,边缘深褐色,病斑可扩大相互愈合成大斑,潮湿时出现很多小黑点.重者病叶率达35～100%,导致植株枯死,病情指数达32～54.笔者1943～1988年,按照方中达(1980)“植病研究方法”进行研究,查对资料,鉴定侵染砂仁引起发病的病原菌是炭疽菌属胶胞     

越南进境人参果炭疽病病原菌的分离鉴定

2008年4月从广州出入境检验检疫局送检的越南进境人参果样品(报检号:P02297)中发现果实表面密布近圆形褐色病斑,病斑直径约为3mm,中央黄褐色,边缘深红褐色,多个病斑可愈合成片.室温下放置1周,病果即完全腐烂,全果果皮及果肉均转成褐色.采用组织分离培养法获得分离物,经对分离物培养性状与形态特征观察、致病性测定,并辅助于分子生物学检测,将病害鉴定为人参果炭疽病,病原菌为胶孢炭疽菌(Colletotrichum gloeosporioides).Blast分析表明从病原菌基因组中扩增到的1.8 kbp 18S rDNA 片段与胶孢炭疽菌18S rRNA基因同源率为99%.迄今为止人参果炭疽病在国内外均未见报道,这是我国首次从进境人参果中截获并检验鉴定该病害.     

新疆茶花上两种叶病新纪录——茶花灰斑病与炭疽病

在新疆茶花上发现了茶花灰斑病和茶花炭疽病两种新病害。茶花灰斑病有两种症状类型 :有限病斑型初期叶上出现黑褐色小点 ,病斑扩大后微凹陷 ,灰褐色至灰白色 ,叶背病斑茶褐色 ,多为不规则形或带圆形 ,病健交界明显 ;无限病斑型多表现在叶缘或叶尖 ,初出现黑褐色小点 ,病斑即速向健康部位呈扇形扩展 ,暗红褐色 ,无边缘 ,其外缘呈墨绿色浸蚀状。茶花炭疽病叶上病斑呈不规则圆形 ,灰白色 ,但近边缘处为深褐色 ,叶背面病斑褐色 ,后期叶部病斑两面同时出现略突起的小黑点物 ,成熟后顶端破裂 ,溢出玫瑰色胶质物。该病与茶花灰斑病的有限病斑类型的症状相似 ,但可视叶背面有无小黑点相区分     

Disease resistance in Vicia faba and Phaseolus vulgaris

Droplets of spore suspensions of each of four isolates ofBotrytis cinerea but not those of each of four isolates ofB. fabae proved to contain an antifungal compound 24 h after application on pod tissue ofVicia faba. Partial inhibition of germ-tube growth of three highly pathogenic isolates ofB. fabae was caused at 2.5 times the concentration of inhibitor needed to cause similar inhibition of each isolate ofB. cinerea and a weakly pathogenic isolate ofB. fabae. After extraction, concentration and chromatographic separation, 5–10 times more inhibitor was obtained from lesions in pods caused byB. cinerea than from those caused byB. fabae. However, the amounts of inhibitor extracted from whole leaves bearing either large lesions caused byB. fabae or small lesions caused byB. cinerea were almost the same. It is suggested that infection by either fungus induces inhibitor formation, but thatB. fabae metabolizes the inhibitor to an inactive form.No relation was found between amounts of an inhibitor produced in droplets of spore suspensions 24 h after application on pods of differential varieties ofPhaseolus vulgaris and the disease reactions caused by races ofColletotrichum lindemuthianum. Each race appeared to have a similar sensitivity to the inhibitor. Anatomical studies showed that only superficial growth of germ-tubes occurred in seed cavities in the first two days, after which penetration took place. Resistant or susceptible reactions were distinguished after 6 days in young pods, and even later in old pods. Before rejecting the hypothesis that the inhibitor may have a role in the mechanism of disease resistance, amounts of inhibitor in and around infection sites on leaves or stems should be measured. Apparent protection of leaf areas against infection was caused by prior inoculation with a race which was avirulent on the leaf. This phenomenon is consistent with the action of an inhibitor of the type found in pod tissues, but could be caused by reactions as yet unknown.     

Real-time quantitative PCR assays for evaluation of soybean varieties for resistance to the stem and root rot pathogen Phytophthora sojae

Phytophthora root and stem rot caused by Phytophthora sojae is one of the most destructive disease of soybeans in the world. Effective management of the disease depends on selection and use of soybean varieties resistant to the disease. Fast and reliable procedures are vital to screen soybean varieties against the pathogen. Novel real-time quantitative (qPCR) assays were developed for both absolute and relative quantification of P. sojae in infected root tissues. QPCR assays were based on the detection of the internal transcribed spacer (ITS) gene of the pathogen and 18S ribosomal gene of the host plant. Absolute qPCR allowed the detection of as low as 10 femtograms (fg) of P. sojae DNA in soybean roots. Relative qPCR, employing the comparative threshold cycle (Ct) method, was effective and reliable for quantification of P. sojae DNA normalized to plant DNA in infected soybean root tissues. P. sojae DNA quantities detected in both qPCR assays had high correlations with disease severity index (DSI) ratings of soybean varieties. QPCR assays developed in this study were useful for determination of the levels of P. sojae DNA in different varieties of soybean and for evaluation of them for relative resistance to the pathogen.     

Trichoderma viride induces phenolics in groundnut (Arachis hypogaea L.) seedlings challenged with rot pathogen (Aspergillus niger Van Tieghem)

The study showed significant differences in percent collar rot disease incidence in groundnut varieties grown in non-infested soil (T₁), challenged with pathogen – Aspergillus niger (T₂), and pathogen + Trichoderma viride 60 (T₃) treatments. Total phenols revealed a significantly higher content in tolerant varieties (J-11, GG-2) of groundnut compared with moderately susceptible (GAUG-10, GG-13) and susceptible (GG-20) varieties. The phenol content accumulated at a higher rate (193%) in GG-20, followed by GG-2 (146%) and J-11 (107%) varieties during disease development stages. HPLC analysis detected six major phenolics, viz., hydroquinone, gallic, chlorogenic, ferulic, salicylic and cinnamic acids. Among six peaks, hydroquinone was found highest in GG-2 at 3 days in T₃. Gallic and salicylic acids increased up to 9 days, while ferulic acid continued to induce up to 15 days in tolerant varieties (J-11, GG-2) of Trichoderma-treated (T₃) seedlings. A correlation study indicated that Trichoderma treatment induced five phenolics – except gallic acid – with a higher level of significance in a susceptible variety to reduce disease incidence compared with tolerant varieties. Results demonstrate the T. viride 60 mediated systemic induction of phenolics for biologic control and their probable role in protecting groundnut against A. niger infection.     

哈茨木霉对黄瓜尖孢镰刀菌的抑制作用和抗性相关基因表达

采用室内筛选与田间防效相结合的方法,对哈茨木霉抑制黄瓜尖孢镰刀菌的拮抗机制进行了研究。对峙培养结果显示,木霉菌和病原菌间均形成了较明显的抑菌圈,对病原菌的抑菌率达66.7%~85.8%,其中菌株TG、TM对枯萎病菌抑制作用较强。木霉菌可有效提高根系对病原菌的抗性,黄瓜植株接种枯萎病菌后,根系细胞大量死亡,而先接种木霉菌再接种病原菌后则减小了对根系的伤害。田间防效试验结果表明,TG和TM孢子悬浮液浓度在108个/mL时防效最好,分别为54.9%和49.4%。接种木霉菌植株根系抗性基因的表达量均高于对照植株,呈双峰趋势。在第6天时抗性基因表达量最高,WRKY6、MYB、PR-1、PAL、GST和GLU的表达量分别为对照的5.15、5.22、6.07、6.00、3.16、和16.15倍。表明木霉菌通过激活与胁迫相关的基因表达提高了对病原菌的抗性。     

Peroxidase activity in rice leaves infected withHelminthosporium oryzae in relation to lesion maturation

Two-week-old seedlings of two rice varieties, resistant CH13 and susceptible Benibhog, were inoculated with a conidial suspension ofHelminthosporium oryzae. The resistant host exhibited greater peroxidase activity than the susceptible one when infected with the pathogen. Enzyme activities were measured 0, 24, 48, 72, 96 and 120 h after infection. Peroxidase activity in diseased leaves bearing young lesions,i.e., 24 h after inoculation, was not appreciably different from that in non-infected leaves. At the intermediate and mature stages of lesion development,ri.e., 72 and 120 h after inoculation, a marked increase in peroxidase activity was observed. The pattern of increased enzyme activity was similar in both varieties but the whole event proceeded faster in the resistant variety. No marked change in peroxidase activity in healthy leaves of both the varieties was noticed with increase of seedling age.     

Inoculation with <Emphasis Type="Italic">Sclerotium rolfsii</Emphasis>, cause of stem rot in Jerusalem artichoke,under field conditions

Stem rot caused by Sclerotium rolfsii is an important problem of Jerusalem artichoke, and breeding of Jerusalem artichoke for resistance to stem rot requires effective screening methods. The objective of this study was to compare methods for inoculating Jerusalem artichoke with S. rolfsii under field conditions. A 4 × 2 × 3 factorial in a randomized complete block with four replications was used in two environments characterized by different rates of fertilizer application (recommended rate and low rate) in the rainy season. The factors included four Jerusalem artichoke varieties (HEL280, HEL278, HEL256 and JA49), two levels of wounding (wounded and not wounded) and three methods of inoculation. The inoculation methods consisted of: 1) non-inoculated natural infection; 2) attaching one colonized sorghum seed at the crown of plants (single sorghum seed method); and 3) spreading 30 g m^?2 of colonized sorghum seeds (broadcast inoculation method). Jerusalem artichoke varieties and inoculation methods were significantly different for disease incidence, whereas the difference between wounded and non wounded treatments was not significant. Significant interactions were found between the variety and wounding method, the variety and inoculation method, wounding method and inoculation method, and inoculation method and environments. Natural infection resulted in the lowest disease incidence (32.2 %), whereas the single sorghum seed and the broadcast inoculation methods had a high disease incidence (79.0 % and 77.3 % respectively) and were not signnificantly different from each other. Broadcast inoculation did not allow differentiation of Jerusalem artichoke varieties for disease incidence, whereas single seed inoculation could better identify the differences among Jerusalem artichoke varieties.     

Incidence of grapevine trunk diseases is influenced by soil,topology and vineyard age,but not by <Emphasis Type="Italic">Diplodia seriata</Emphasis> infection rate in the Tokaj Wine Region,Hungary

Grapevine Trunk Diseases (GTD) are of great importance worldwide, including Hungary, a Center European country with long wine producing history. Several GTD pathogens have been described till now in Europe, but only a few from Hungary. The presence of a GTD pathogen in the vine does not necessarily result in the immediate appearance of disease symptoms, and information on the importance of environmental factors related to disease incidence are still limited. The aim of this research was to assess the occurrence of GTD in the Tokaj Wine Region, and to determine the biotic and abiotic factors influencing disease incidence. Five vineyards within 15 km radius – each with different topology, soil types, varieties and age – were studied for three consecutive years (2013 – 2015). The incidence of GTD-infection was determined every year for each vineyard. Diplodia seriata was isolated with incidence ranging from 50 to 100%, while Diaporthe spp. were the only other – minor – GTD pathogen found. Topology and soil type appeared to be major abiotic factors affecting incidence of GTD symptom. Disease incidence was also positively correlating with the age of the vineyards, and it was in fact found to be the definitive biotic factor regarding incidence. In contrast, D. seriata infection rate appeared unrelated to disease incidence or to any of the biotic or abiotic factors investigated.     

水稻稻曲病室内人工接种技术

为提高稻曲病人工接种的发病效果和稳定性,在温棚条件下采用水稻孕穗期注射接种法分别研究了稻曲病菌不同接种体、培养时间、接种浓度和接种时期的接种效果。采用病菌马铃薯蔗糖液体培养基（potato sucrose broth,PSB）作为接种体,其穗发病率为100%,明显好于病菌米糠培养液（23.33%）。病菌在PSB中培养5～7天接种效果较好,随病菌培养天数的延长,接种效果明显下降。接种的分生孢子浓度越低水稻病穗率和病粒数也越低。在水稻品种两优培九孕穗中后期,采用含分生孢子浓度为4×106个/mL的病菌PSB培养液注射接种,穗发病率达100%,平均病粒数35.1粒,最高达87粒。研究表明,温棚条件下建立的稻曲病人工接种技术能获得稳定的发病效果,并可区别水稻品种间的抗性差异。     

Evaluation of oilseed rape seed yield losses caused by <Emphasis Type="Italic">Leptosphaeria biglobosa</Emphasis> in central China

Phoma stem canker of oilseed rape (Brassica napus), caused by Leptosphaeria maculans/L. biglobosa is a globally important disease. Severe phoma stem canker symptoms have been observed on winter oilseed rape in China but the seed yield loss caused by this disease remains unknown. In May 2012 and May 2013, 17 and 13 crops were surveyed, respectively, in seven counties of Hubei Province, central China. Stems with phoma stem canker disease symptoms were sampled for pathogen isolation and identification. Only L. biglobosa was identified by culture morphology and species-specific PCR; no L. maculans was found. To evaluate the yield losses, yield components (number of branches per plant, number of pods per plant, 1000-seed weight, number of seeds per pod) were assessed on healthy and diseased plants sampled from crops in four counties and on plants from inoculated pot experiments (plants of three cultivars were inoculated at the green bud stage by injecting L. biglobosa conidia into the stem between the first and second leaf scars). Results of the field surveys showed that diseased plants had 14–61% less branches and 32–83% less pods than healthy plants, respectively. The estimated seed yield loss varied from 10% to 21% and from 13% to 37% in 2012 and 2013, respectively. In the pot experiments, there were no differences in numbers of branches or pods but there were differences in number of seeds per pod between inoculated and control plants. For the three cultivars tested, the inoculated plants had yield losses of 29–56% compared with the control. This study indicates that L. biglobosa could cause substantial seed yield loss in China.